An RNA-Cleaving DNAzyme That Requires an Organic Solvent to Function |
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Authors: | Dr Tianjun Chang Guangping Li Dingran Chang Ryan Amini Xiaoni Zhu Prof Tongqian Zhao Jimmy Gu Prof Zhongping Li Prof Yingfu Li |
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Institution: | 1. School of Resources and Environment, Henan Polytechnic University, Jiaozuo, 454000 China;2. Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, L8S 4K1 Canada;3. Institute of Environmental Science, Shanxi University, Taiyuan, 030006 China |
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Abstract: | Engineering functional nucleic acids that are active under unusual conditions will not only reveal their hidden abilities but also lay the groundwork for pursuing them for unique applications. Although many DNAzymes have been derived to catalyze diverse chemical reactions in aqueous solutions, no prior study has been set up to purposely derive DNAzymes that require an organic solvent to function. Herein, we utilized in vitro selection to isolate RNA-cleaving DNAzymes from a random-sequence DNA pool that were “compelled” to accept 35 % dimethyl sulfoxide (DMSO) as a cosolvent, via counter selection in a purely aqueous solution followed by positive selection in the same solution containing 35 % DMSO. This experiment led to the discovery of a new DNAzyme that requires 35 % DMSO for its catalytic activity and exhibits drastically reduced activity without DMSO. This DNAzyme also requires divalent metal ions for catalysis, and its activity is enhanced by monovalent ions. A minimized, more efficient DNAzyme was also derived. This work demonstrates that highly functional, organic solvent-dependent DNAzymes can be isolated from random-sequence DNA libraries via forced in vitro selection, thus expanding the capability and potential utility of catalytic DNA. |
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Keywords: | DNAzyme Dimethyl Sulfoxide Functional Nucleic Acid In Vitro Selection Organic Solvent |
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